A Glycosidic-Bond-Based Mass-Spectrometry-Cleavable Cross-linker Enables In Vivo Cross-linking for Protein Complex
Jing Chen1,2, Qun Zhao1, Hang Gao1,3
1CAS Key Labratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, 116023, China.
Angewandte Chemie (International Ed. in English)
|March 30, 2023
Summary
A novel MS-cleavable cross-linker, trehalose disuccinimidyl ester (TDS), enhances in vivo chemical cross-linking mass spectrometry (CXMS) by simplifying data analysis and improving biocompatibility for protein complex studies.
Area of Science:
- Biochemistry
- Proteomics
- Chemical Biology
Background:
- Chemical cross-linking mass spectrometry (CXMS) is vital for protein complex analysis.
- In vivo CXMS is hindered by cross-linker biocompatibility and complex data analysis.
Purpose of the Study:
- To develop a biocompatible and MS-cleavable cross-linker for enhanced in vivo CXMS.
- To improve the accuracy and throughput of protein complex identification in living systems.
Main Methods:
- Design and synthesis of a glycosidic bond-based MS-cleavable cross-linker: trehalose disuccinimidyl ester (TDS).
- Fragmentation of TDS in MS under CID/HCD to yield single peptides.
- Evaluation of TDS cell-penetrating properties and water solubility.
Main Results:
- TDS simplifies cross-linked peptides into single peptides via selective MS cleavage.
- Significantly enhanced cross-linking identification accuracy and throughput.
- Demonstrated cell-penetrating properties and high water solubility, reducing DMSO dependence.
Conclusions:
- TDS offers a biocompatible and accurate toolkit for in vivo CXMS.
- The MS-cleavable nature of TDS streamlines data analysis for protein complex characterization.
- TDS facilitates advanced CXMS studies in living systems.
Keywords:
Cleavage ReactionsGlycosidicsMass SpectrometryProtein StructuresProtein-Protein Interactions
